FIG 4.

RNF31 is cleaved at aspartates 348, 387, and 390. (A) WB analysis of 293T cells transfected with RNF31 tagged with Myc at the N terminus, followed by treatment with TNF-α (40 ng/ml) and CHX (10 μg/ml). (B) Estimated sites of RNF31 cleavage by caspases. (C) WB analysis of 293T cells transfected with a plasmid encoding Myc-conjugated WT, D390A, D348/390A, or D348/387/390A RNF31. (D) Results of an in vitro cleavage assay in which recombinant WT or D348/387/390A mutant RNF31 was incubated with or without caspase 8, caspase 3, or caspase 6 for 1 h.